DAG: Live Cell Diacylglycerol Assays
Bright Fluorescent Biosensors for Diacylglycerol
- Genetically-encoded for low toxicity
- Plate reader or imaging system compatible
- Choose red or green fluorescence
Downward DAG starts out bright and decreases in fluorescence when diacylglycerol levels increase. This assay is easily detected on fluorescence plate readers and helps with imaging cells that have low expression efficiency.
Upward DAG starts out dim and increases in fluorescence when when diacylglycerol levels increase. This DAG assay is easily detected on fluorescence plate readers or imaging systems.
DAG Assay Performance
Off the shelf kits include the DAG sensor in BacMam, a BSL-1 viral vector for efficient delivery to most cell types. Purified BacMam, AAV, Lenti vectors by request.
- Easily detectable on plate readers or imaging systems
- Consistent well-to-well expression
- High signal-to-noise ratio
Red DAG sensor multiplexed with a green cAMP sensor indicates dose-dependent signaling via a calcitonin receptor.
In HEK 293 cells over-expressing the P2Y11 receptor, ATP produces changes in both DAG and Ca2+, consistent with activation of Gq-coupled GPCR signaling. UTP produces no DAG response, indicating that the Ca2+ change is independent of Gq.
Multiplex Assays
- Combine red and green sensors to measure multiple GPCR pathways simultaneously
- Red or green sensor version
- Single channel, non-FRET signal
A Robust & Versatile DAG Assay Kit
- Detect diacylglycerol increases in real time
- Screen Gq coupled GPCRs in living cells
- Viral vector delivery to practically any cell type
- Target to microdomains or specific cells in mixed cultures
Simple Protocol
- Minimal liquid handling
- No cell lysis
- 96 or 384 well compatible
Recent Publications
- C Deisl et al. Hypertrophy of human embryonic stem cell–derived cardiomyocytes supported by positive feedback between Ca2+ and diacylglycerol signals. Pflügers Archiv – European Journal of Physiology Jun. 2019
- Herranz, Gonzalo et al. Protein Kinase C δ Regulates the Depletion of Actin at the Immunological Synapse Required for Polarized Exosome Secretion by T Cells. Frontiers in Immunology Apr. 2019
- Ohno, et al.Dynamics of Presynaptic Diacylglycerol in a Sensory Neuron Encode Differences between Past and Current Stimulus Intensity. Cell Reports, Vol. 20, Issue 10. Sept. 2017
- Shigeto, et al. GLP-1 stimulates insulin secretion by PKC-dependent TRPM4 and TRPM5 activation.J Clinical Invest. 2015. doi:10.1172/JCI81975.
- P. Rorsman, et al. GLP-1 stimulates insulin secretion by PKC-dependent TRPM4 and TRPM5 activation J Clinical Investigation Nov. 2015
- Arttamangkul, et al. Does PKC activation increase the homologous desensitization of μ opioid receptor? British Journal of Pharmacology.Jan. 2015.
- Xingjuan, et al. PKC-dependent Phosphorylation of the H1 Histamine Receptor Modulates TRPC6 Activity.Cells. 2014.
- P. Tewson, et al. New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories. J Biomol Screen Dec. 2015. Request Full Text PDF
- J. Williams, et al. Does PKC activation increase the homologous desensitization of μ opioid receptors? British Journal of Pharmacology April 2014
- Tewson, et al. Simultaneous Detection of Ca2+and Diacylglycerol Signaling in Living Cells.PLoS One. 2012.
- C. Peters, et al. Simultaneous detection of GPCR second messengers in living cells. BMG LabTech Dec. 2016.
“Assay is working beautifully with cardiomyocytes, showing a strong and immediate response.”
Expression in human islets is pretty superb, after 18 hours of incubation. An excellent probe!
